Olgun F Ayca Ozdemir, Ozyurt Dilek, Berker Kadriye Isil, Demirata Birsen, Apak Resat
J Sci Food Agric. 2014 Sep;94(12):2401-8. doi: 10.1002/jsfa.6569.
This study proposes modifications to the conventional Folin-Ciocalteu (FC) spectrophotometric method for individually determining ascorbic acid (AA) in complex matrices in the presence of other phenolics and potential interferents. The conventional FC assay in the aqueous phase,which normally measures total water-soluble phenolics and other antioxidants, has recently been modified by incorporating isobutanol (iso-BuOH) in the solvent mixture for the simultaneous determination of lipophilic and hydrophilic antioxidants in foods.
Interference effects of other flavonoids and phenolics to individual AA assays were overcome by preliminary extraction–removal as their La(III) chelates into ethyl acetate (EtAc). The pH of the medium was adjusted to 3.0 in order to prevent conversion of AA into over-oxidation products further beyond the dehydroascorbic acid stage, as encountered in the conventional FC assay carried out at pH 10. This pH does not permit most phenolics to ionise, rendering their oxidation difficult.
Both methods (conventional and iso-BuOH-modified FC at pH 3, with and without La(III)/EtAc pre-extraction) were applied to the determination of ascorbic acid in pharmaceutical tablets and orange juice, showing good agreement with HPLC results. The proposed spectrophotometric methods with their low cost, simplicity, reliability, versatility and accuracy offer novelty to the determination of AA in complex matrices.
本研究对传统的福林 - 西奥尔特(FC)分光光度法提出了改进,用于在存在其他酚类物质和潜在干扰物的复杂基质中单独测定抗坏血酸(AA)。水相中的传统FC测定法通常测量总水溶性酚类物质和其他抗氧化剂,最近通过在溶剂混合物中加入异丁醇(iso - BuOH)进行了改进,以同时测定食品中的亲脂性和亲水性抗氧化剂。
通过初步萃取 - 将其他黄酮类化合物和酚类物质作为其镧(III)螯合物去除到乙酸乙酯(EtAc)中,克服了其他黄酮类化合物和酚类物质对单个AA测定的干扰作用。将介质的pH值调节至3.0,以防止AA转化为超出脱氢抗坏血酸阶段的过度氧化产物,这在pH 10下进行的传统FC测定中会遇到。此pH值不允许大多数酚类物质离子化,使其难以氧化。
两种方法(传统的和在pH 3下用异丁醇改性的FC,有和没有镧(III)/乙酸乙酯预萃取)都应用于药物片剂和橙汁中抗坏血酸的测定,结果与高效液相色谱法(HPLC)结果吻合良好。所提出的分光光度法具有低成本、简单、可靠、多功能和准确的特点,为复杂基质中AA的测定提供了新颖性。